Ziehl-Neelsen Staining Method Research Articles

Application of Ziehl Neelsen Staining Method for Taenia spp. Eggs Differentiation.

Three of Taenia species, named Taenia saginata, T. solium and T. asiatica can be found in Indonesia, but only T. solium can lead to neurocysticercosis. The morphology of those 3 Taenia spp. egg is indistinguishable by standard parasitology procedure. We aimed to use Ziehl Neelsen staining for differentiation of eggs of T. saginata and T. solium. As many as 40 containers of stool samples from the positive helminthiasis patients in Jakarta, Indonesia were collected during the year 2018. From each container, 10 slides prepared for staining with Kato-Katz technique as the preliminary examination. From those stool samples with positive taeniasis, we then once again made 10 slides/container for Ziehl Neelsen staining. The first 400 slides stained with Kato-Katz technique as preliminary test were all positive for Taenia spp. The second 400 slides, we got 244 slides that gave good results as we could distinguished between the eggs of T. saginata and T. solium, meanwhile the remainder 156 slides gave unconfirmed results. From those 244 slides, 154 slides showed T. saginata eggs with magenta colored and 90 slides showed T. solium eggs with blue/purple colored. The eggs of T. solium slightly smaller if compared to Taenia saginata and had round shape, meanwhile T. saginata eggs were oval in shape. Ziehl Neelsen staining method can be used as an alternative parasitological method to differentiate the eggs of T. saginata and T. solium.

Commonness and Risk Factors Associated with Gastrointestinal Parasitic Infections among Inmates of Port Harcourt Children’s Home

Background: Gastrointestinal parasites are important health problems among most societies.
 Objectives: Prevalence and risk factors associated with gastrointestinal parasites among inmates of Port Harcourt children home.
 Methods: This prospective study was carried out in August and September 2019 at the Port Harcourt children home. Fecal samples were collected from 63 healthy looking inmates and examined macroscopically and microscopically (using normal saline and iodine wet mount, formalin-ether sedimentation and modified Ziehl Neelsen staining) methods. Finally, data was analyzed using descriptive statistics and Chi-square (X2) test.
 Results: Result showed that 74.6% (47/63) of the female and 25.4% (16/63) of the male tested positive for GIPs. Age group 11-15 years had prevalence of 17(44.8%). The common parasites identified were A. lumbricoides 10(15.9%), T. trichiura 7(11.1%) and hookworm (Ancylostoma duodenale) 2(3.2%). The educational level prevalence of 10(52.6%) was observed among the primary level (p<0.05). The risk factors such as: the use of drug in the last three months, hand washing with soap after using the toilet and washing of hands after contact with soil had significance of p<0.05 while putting on foot wear outside the house p>0.05.
 Conclusion: The distribution of gastrointestinal parasitic infections is high among the children. Necessary treatment intervention safety measures should be routinely carried out to reduce the spread to zero tolerance.

Diagnostic Yield of Xpert MTB/RIF Assay Using Bronchoalveolar Lavage Fluid in Detecting Mycobacterium tuberculosis among the Sputum-Scarce Suspected Pulmonary TB Patients.

Tuberculosis (TB) remains one of the leading causes of death worldwide and is caused by the single infectious agent Mycobacterium tuberculosis (Mtb). Although sputum is the most common specimen for pulmonary TB detection, some other respiratory specimens, such as bronchoalveolar lavage (BAL) fluid, gastric lavage (GL), and induced sputum (IS), are also collected from patients who are unable to deliver sputum. In this study, we aimed to evaluate the diagnostic performances of different test methods for TB diagnosis using BAL fluid specimens from sputum-scarce pulmonary TB patients. In this current study, a total of 210 BAL fluid specimens were collected and subjected to culture on Lowenstein–Jensen (L-J) medium, using an N-acetyl-L-cysteine-Sodium Hydroxide decontamination and digestion method, Xpert MTB/RIF (Xpert, Cepheid, Sunnyvale, CA, USA) assay, and acid-fast bacilli (AFB) microscopy with a Ziehl–Neelsen staining method for the detection of pulmonary TB. The sensitivity and specificity of these methods were then analyzed against the composite reference standard (CRS). Additionally, the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of these assays. Among the 210 specimens, 39 (18.6%), 27 (12.8%), and 12 (5.7%) were found positive with Xpert assay, culture, and AFB microscopy, respectively. Considering the CRS, 42 (20%) were positive as the final diagnosis. The Xpert assay had a significantly higher sensitivity (92.9%, 95% CI: 80.5–98.5) compared to culture (64.3%, 95% CI: 48.0–78.4) and AFB microscopy (28.6%, 95% CI: 15.7–44.6) against the CRS. Additionally, the area under the ROC curve (AUC) for the Xpert assay, culture, and AFB microscopy accounted for 0.964, 0.821, and 0.655, respectively, when using CRS as the reference. In conclusion, our study findings demonstrated that the Xpert assay conferred a considerable diagnostic potential compared to other conventional methods for the diagnosis of pulmonary TB from BAL fluid specimens.

Analysis of The Effectiveness of Mycobacterium Tuberculosis Through Resistant Bacterial Staining and Molecular Rapid Tests

The diagnosis of TB can be confirmed by microscopic examination of smear on the sputum of the patient. However, because this method requires very expensive equipment, it is difficult to implement it in health facilities with simple facilities. Therefore, the Ziehl Neelsen staining method is a fairly simple method of choice and provides high sensitivity and specificity. In the research of Eka Kurniwan, et al. 2016, from the results of examining 40 samples with the Molecular Rapid Test (TCM), 16 people (40%) were found to be positive. The research objective was to know Mycobacterium tuberculosis through sputum examination through Ziehl Nelssen's acid-resistant bacteria staining and Molecular Rapid Test (TCM) of Puskesmas Dempo Palembang based on volume, odor, color and consistency. This type of research is a diagnostic test with a cross sectional study approach. Population and sample of 85 patients. The sampling technique in this study is using the total sampling method. Conclusion There is no effect based on volume, there is an effect based on the smell, color and consistency of sputum, there are differences in the results of Mycobacterium tuberculosis through sputum examination through Ziehl Nelssen's Acid-Resistant Bacteria Staining and Molecular Rapid Test (TCM) at Puskesmas Dempo Palembang.

Risk Factors and Prevalence of Taeniasis among Displaced peoples in Ombadda Omdurman State-Sudan

Background: Taeniasis is infection of human with intestinal cestode tape wormparasites belonged to the genus Taenia species saginata and solium. Objective: The study aimed to determine the risk factors and prevalence of taeniasis among displaced peoples district in Ombadda Omdurman state- Sudan. Material and Methods: This is a cross sectional study was conducted in Ombadda and Basheer hospitals in Omdurman-Sudan from the period September to November2021.A total of 150 stools samples were collected and examined using direct microscopy, formal ether concentration technique and Ziehl-Neelsen staining method fordetecting TaeniaSpp. Results: A total of 150stools samples collected and examined, 10 (6.3%) out of 150 faecal samples were positive and 140 (93.7%) were negative sample. Out of 10 positive samples 6 (60%) were males and 4 (40%) were females their age groups range from 20 – 60 Years old, The Eggs stage of Taenia saginata and Taenia solium was identified by using direct microscopy and formal ether concentration t techniques and differentiated by the Ziehl-Neelson staining method.

Gastrointestinal parasites in immunocompromised patients; A comparative cross-sectional study

Gastrointestinal parasites (GIPs), including helminths and protozoa species, are a major health problem in many parts of the world. About 3.5 billion people are affected by the parasites worldwide. GIPs are one of the leading causes of death among immunocompromised individuals and can cause serious clinical complications, especially in people with Human Immunodeficiency Virus (HIV)/AIDS, hemodialysis patients, and transplant recipients. This study aimed to compare the prevalence of GIPs among immunocompromised patients and immunocompetent individuals in Lorestan province, West Iran. In the current study, with a sampling of 232 participants (114 hemodialysis, AIDS, and organ transplantation immunocompromised patients and 118 immunocompetent individuals as the control group), demographic characteristics and risk factors for GIPs were collected through a pre-designed questionnaire. Stool samples of patients and the control group were examined for GIPs using different diagnostic methods including direct smear (saline and Lugol's iodine), Ziehl-Neelsen staining, agar-plate culture, and concentration method (formalin ether sedimentation). To evaluate the relative status of the immune system, TCD4+ cells were counted in the blood samples of the subjects by flow cytometry. The results were analyzed using SPSS 21 software, Fisher exact, and chi-square statistical tests. Multivariate modeling of the data was performed using logistic regression. The prevalence of GIPs in immunocompromised patients was more than twice that of immunocompetent individuals in the control group (42.06% vs. 17.79%). The most prevalent parasites identified among immunocompromised patients were Cryptosporidium sp. (27.1%), Blastocystis sp. (16.7%), and Entamoeba coli (14.6%) respectively. Cryptosporidium sp. had the highest frequency among hemodialysis patients (6.49%), AIDS patients (26.92%), and transplant recipients (18.18%) respectively. Patients with AIDS had the highest positive results for Cryptosporidium sp. followed by Microsporidia sp. (23.7%). In immunocompetent individuals, the highest prevalence of GIPs was related to Blastocystis sp and Trichomonas hominis (28.57%). Statistical analysis of the data showed that there was a statistically significant difference between various age groups regarding infection with GIPs so the highest rate of GIPs infection was observed in the age group lower than 50 years (P = 0.035). The statistical difference between the variable of location and infection with GIPs was insignificant but remarkable (P = 0.070). According to the results, it can be concluded that GIP is more common in immunocompromised patients than in immunocompetent individuals with cryptosporidium sp. predominance. Due to the favorable conditions of immunocompromised patients for GIPs and considering them as one of the important sources of parasitic infections and parasite transmission in society, control, prevention, and monitoring of their social behaviors along with health issues are inevitable.

Epidemiology of cryptosporidiosis in dairy calves in central and eastern Algeria

The purpose of our survey is to estimate the frequency of Cryptosporidium spp in dairy calves under 60 days of age, the pathogenic role of Cryptosporidium spp in the development of diarrhea, the intervention and association of other enteropathogens often involved in neonatal diarrhea and the possible relationship between infection and certain farming practices in order to estimate the share of risk factors, and on the other hand, to compare between the modified Ziehl-Neelsen staining method and the copro-ELISA test. We found 35 positive samples out of 223 faecal samples analyzed by modified Ziehl-Nielson staining, with a prevalence of 15.69%, the infected calves had 4 times the risk of diarrhea (RR= 4), the age group [8-15 days] is the most susceptible to infection compared to the other age groups (31.1%; p<0.05). The ELISA test revealed that 33 out of 92 calves were infected by Cryptosporidium spp., with a prevalence of 35.87%, showing a sensitivity and specificity of Ziehl-Neelsen staining of 51.5% and 96% respectively.

Intestinal Parasitosis among Human Immunodeficiency Virus and Tuberculosis Infected Patients from Dharan, Nepal

Intestinal parasitic infestation reflects a health threat with patients living with HIV & Tu-berculosis (TB). Parasitic infections caused by protozoa and helminths are the most com-mon infections worldwide. The present study was done to find out the frequency of intes-tinal parasitosis among HIV and Tuberculosis (TB) patients of Dharan total of 53 samples (>15 years) from HIV seropositive patients and 28 samples (>20years) from TB patients were collected in a clean, dry and capped fitted container and subjected to macroscopic and microscopic examination for ova, cyst, adult parasites and or segments of parasites. Stool samples were fixed in a 10% formalin-ether solution. Sedimentation technique, with modified acid-fast (Ziehl-Neelsen) staining method, was performed for opportunistic in-testinal parasites in both patients. Multi-parasitic infection was noted in the study. The overall prevalence of intestinal parasites was found to be 54 (66.67%). This result had shown that the participation of male patients was comparatively higher than female pa-tients. Out of 81 stool samples, 15 (27.78%) G. lamblia, 12(22.22%) E. histolytica, 12 (22.22%) Cryptosporidium parvum, 6 (11.11%), Isospora belli 4 (7.40%) Microspridium, 1(1.85%) Blastocystis hominis, 2(3.70%) and Taenia spp, 2(3.70%). To prevent this infec-tion appropriate health education should be given to the patients concerning disease trans-mission, antiparasitic therapy, personal hygiene, and safe drinking water.

Effects of thymulin 5cH in granuloma evolution and B1 cell differentiation: an experimental model to understand its biological mechanisms

In previous studies, we found that thymulin (a thymic hormone), when prepared in homeopathic 5cH potency, had the property to improve the productive performance of broiler chickens infected with reovirus, as well as modulate the development of Ehrlich tumor and granuloma inflammatory lesions in mice by immune-mediated mechanisms. The aim of the present work was to study the immunomodulatory mechanisms of thymulin 5cH in a granuloma experimental model, by subcutaneous inoculation of BCG in mice, focusing the B-1 cells and zinc involvement in this process. Three groups of male Balb/c SPF mice (group A treated with thymulin 5cH, group B treated with thymulin 5cH incubated in Chelex ® - a zinc chelant - and group C, control, treated with vehicle) were inoculated with BCG in the left footpad and subcutaneous granuloma and spleen were harvested for histomorphometry analysis, after 7, 14 and 21 days. Ziehl-Neelsen, HE and Prussia Blue staining methods were used. Flow cytometry was also used in the same times to characterize and quantify peritoneal cells. Positive cells for CD11b (activated phagocytes, B-1 cells), CD19 (B-1 and B-2 cells), CD23 (negative B-1 cells, positive B2 cells) and CD5 (B-1a cells) were analyzed in a FACS Calibur (BD) device. Statistical analysis was performed using Kruskal - Wallis / Dunn for nonparametric evaluations and ANOVA / Tuckey-Krammer for the parametric ones. The X² method was used to evaluate the cell count in flow cytometry. P values ≤ 0.05 were considered statistically significant. Mice treated with thymulin 5cH presented higher macrophage activity and increase in the follicular area were seen in spleen after 7 days. Increase in gross lesion diameter and decrease in local BCG infection were seen after 21 days. At this time, the flow cytometry demonstrated the increase in peritoneal phagocytes derived from B-1 cells in thymulin 5cH treated mice, independently of Chelex ® incubation. The incubation of thymulin 5cH with Chelex ® blocked its effects only upon the number of B2 cells in the peritoneum and reduces Mn levels in the medicine solution. We conclude that thymulin 5CH modulates the BCG-induced granuloma through more than one mechanism, especially by peritoneal B1 cell differentiation into phagocytes.

A Mycobacterium species for Crohn's disease?

In ruminants Mycobacterium avium subspecies paratuberculosis (MAP) is the causative organism of a chronic granulomatous inflammatory bowel disease called Johne's disease (JD). Some researchers have hypothesised that MAP is also associated with Crohn's disease (CD), an inflammatory bowel disease in humans that shares some histological features of JD. Despite numerous attempts to demonstrate causality by researchers, direct microbiological evidence of MAP involvement in CD remains elusive. Importantly, it has not been possible to reliably and reproducibly demonstrate mycobacteria in the tissue of CD patients. Past attempts to visualise mycobacteria in tissue may have been hampered by the use of stains optimised for Mycobacterium tuberculosis complex (MTB) and the lack of reliable bacteriological culture media for both non-tuberculous mycobacteria (NTM) and cell-wall-deficient mycobacteria (CWDM). Here we describe a Ziehl-Neelsen (ZN) staining method for the demonstration of CWDM in resected tissue from patients with Crohn's disease, revealing the association of CWDM in situ with host tissue reactions, and posit this as a cause of the tissue inflammation. Using the ZN stain described we demonstrated the presence of CWDM in 18 out of 18 excised tissue samples from patients diagnosed as having Crohn's disease, and in zero samples out of 15 non-inflammatory bowel disease controls.

Investigation of the relationship between lymphocyte subsets and intestinal parasites

We analyzed the peripheral blood lymphocyte subsets of cancer patients infected with intestinal parasites, with an aim to find out the relationship between the levels of different types of lymphocytes with the prognosis of patients. 201 cancer patients aged 18 and over were included. Stool samples of the patients were examined using native-lugol, trichrome, modified trichrome (Weber's Trichrome stain), and modified Ziehl-Neelsen staining methods. Microsporidia and Cryptosporidium parvum were investigated at the genus and species levels using PCR. Lymphocyte subsets were determined by flow cytometry in blood samples. One or more parasite species were detected in 115 (56.7%) patients. The most common parasite species were Microsporidia, Blastocystis and Entamoeba coli, respectively. The frequency of parasites was high in patients with low lymphocyte percentage, CD3+ T cell and CD3+ CD4+ T (Th) cell levels in blood samples studied by flow cytometry. Microsporidia infection was significantly higher in patients with low lymphocyte percentage and Th cell levels. Similarly, C. parvum infection was found to be significantly higher in patients with low T lymphocyte percentage and Th cell level. Finally, Blastocystis infection was significantly higher in patients with low lymphocyte percentage and CD4/CD8 ratio higher than 1. The decrease in lymphocyte percentage, CD3+ T cell and Th cell count, and low CD4/CD8 ratio in cancer patients increase the frequency of intestinal parasitic infections. Based on these results, lymphocyte subsets may help identify cancer patients at high risk of opportunistic parasites. We suggest that opportunistic parasitic infections affecting the clinical course of the disease should be considered by clinicians during the follow-up and treatment of patients.

Fluorescein Diacetate Vital staining for detecting viability of acid-fast bacilli in sputum of pulmonary tuberculosis patients

ObjectiveTo compare the performance of the Fluorescein diacetate (FDA) vital staining method with Ziehl-Neelsen staining method in detecting the viability of acid-fast bacilli using MGIT culture as “reference standard”. MethodsThis was a single centre prospective observational study conducted from October 2015 to November 2016. Microbiologically confirmed ZN-Smear positive (3+) sputum specimens were obtained from 30 pulmonary tuberculosis patients taking anti-tuberculosis treatment at DOTS centre of NITRD, New Delhi. Patients were made available to collect the first baseline sputum sample before commencing treatment, and an early morning sputum sample was collected as per RNTCP guidelines. After starting treatment, sputum specimens were collected weekly in the first month and thereafter twice-weekly until 18th week. All sputum specimens from patients receiving anti-tuberculosis treatment were examined using Ziehl-Neelsen (ZN) smear microscopy, FDA vital staining, and MGIT culture. ResultOut of 360 follow up sputum specimens collected from 30 adult microbiologically confirmed ZN- Smear (3+) pulmonary tuberculosis patients, 146 were ZN-positive and 130 FDA-positive. Of 130 FDA-positive sputum samples, mycobacteria tuberculosis (MTB) growth was found in 116 sputum samples, of which 116 sputum specimens were positive for FDA. Additionally, 14 culture-negative specimens were FDA positive. No FDA-negative sputum samples were positive for MGIT culture. Among ZN positive specimens, FDA had 100% sensitivity and 85.3% specificity with an accuracy of 96.58% for the detection of viable mycobacteria. Among ZN negative sputum specimens, FDA had comparatively high specificity (95.7%). Using positive MGIT culture as a reference for viability, negative predictive value (NPV) and positive predictive value (PPV) from FDA vital staining method were found to be 100 and 89% respectively. ConclusionFDA staining is a simple and rapid tool for identifying viable MTB bacilli. Because of its excellent NPV and encouraging specificity, FDA staining is useful to identify patients with non-viable bacilli (FDA negative) among retreatment cases at diagnosis and patients on anti-tuberculosis treatment for both drug-sensitive and drug-resistant tuberculosis for follow up for the response of treatment.

Comparison of Fine Needle Aspiration Cytology, Ziehl-Neelsen Staining and Gene X-pert Methods in Suspected Cases of Tubercular Lymphadenopathy

Comparison of Fine Needle Aspiration Cytology, Ziehl-Neelsen Staining and Gene X-pert Methods in Suspected Cases of Tubercular Lymphadenopathy

Correlation of bacterial index to zinc serum level in multibacillary type leprosy patient

Background: Leprosy Multibacillary type is the leading source of infection because total of bacteria is high. Zinc is an essential trace element of viability of host or pathogen. This research aimed to determine the relationship between bacterial index with zinc serum level in patients with leprosy multibacillary type.Methods: It was cross-sectional observational analytic research in 31 subjects selected using consecutive sampling and they met inclusion and exclusion criteria conducted in Outpatient Installation at RSUD Dr. Saiful Anwar Malang and RS Kusta Kediri in February to March 2019. Examination of acid-resistant bacteria used Ziehl Neelsen's staining method and measurement based on Ridley and Joplin. Test/examination of zinc serum level was conducted in Prodia Laboratory using Inductively Coupled Plasma-Mass Spectrometer (ICP-MS). data were analyzed statistically using program of Statistical Package for Social Sciences (SPSS) version 24.Results: From 31 research subjects, it was obtained that the lowest bacteria index of 1+ and the highest bacteria index of 6+ with median value of 3.5±2. the average of zinc serum level was 61.81±13.28 µg/dL with the lowest level of 37.00 µg/dL and the highest of 94.00 µg/dL. Based on Spearman correlation test, it was obtained correlation value of - 0.557 (55.7%) and value p = 0.001.Conclusion: There is a significant correlation between bacteria index and zinc serum level with strong closeness value. Correlation is negative, the higher bacteria index, the lower zinc serum level.

Molecular Detection of Brucella melitensis Associated Reproductive Failure in Native Sheep Flock: Potential Public Health Pathogen from Tropical Region of Tamil Nadu

Brucellosis remains a global burden since the last century. In India, both animal and human brucella infections drastically increase every year. Outbreaks of 40 ewe’s abortion were investigated in native sheep’s from the tropical region of Tamil Nadu. Aborted maternal and fetal tissues were initially subjected to Stamps Modified Ziehl Neelsen (S-ZN) staining method. The DNA was extracted and screened for Brucella genome by Brucella cell surface salt extractable proteins (bcsp31) gene based PCR assay. Further, PCR positive samples were subjected for AMOS PCR for B. abortus, B. ovis, B. melitensis and B. suis identification and differentiation. S-ZN staining of aborted tissues revealed pink, coccobacilli organisms giving presumptive identification of Brucella sp. Molecular detection of brucellosis by bcsp31 gene based PCR in placental tissues and fetal abomasal contents of both the animals yielded a specific amplicon of 223 bp confirming it as Brucella genus infection. Molecular detection and differentiation by AMOS PCR revealed a specific amplicon of 731 bp indicating the involvement of B. meletensis infection in the native free-ranging sheep flock. It is a potent public health pathogen, highly essential to be monitored for the strategic control of brucellosis.

Comparison of Fine Needle Aspiration Cytology, Ziehl-Neelsen Staining and GeneXpert Methods in Suspected Cases of Tubercular Lymphadenopathy

Introduction: The Extrapulmonary Tuberculosis (EPTB) is challenging due to the pauci-bacillary nature of the disease and limited tests available for early diagnosis. It is the most common extrapulmonary manifestation of tuberculosis. The samples from extrapulmonary infection will have low bacterial count as compared with sputum specimens. It is also necessary to rule out other causes of granulomatous inflammation on Fine Needle Aspiration Cytology (FNAC) and confirmation by Ziehl Neelsen (ZN) stain and Cartridge Based Nucleic Acid Amplification Test (CBNAAT). In December 2010, World Health Organisation (WHO) recommended GeneXpert/CBNAAT to be used as the initial diagnostic test in suspected EPTB cases. Aim: To compare FNAC and ZN stain with CBNAAT in the diagnosis of suspected tubercular lymphadenopathy in a tertiary care centre and to know the importance of CBNAAT in the modern era. Materials and Methods: This was a hospital based prospective study carried out over a period of 12 months (July 2019 to June 2020) in the Department of Pathology and Microbiology, ESIC Medical College. All presumptive cases of tubercular lymphadenopathy and purulent aspirates from the lymph nodes of various sites were included in the study. Smears were made after FNA and stained with Haematoxylin and Eosin (H&E) stain and ZN stain and sample was also processed for CBNAAT in all cases of lymphadenopathy. Statistics were done using SPSS software version 20.00. Results: The total number of cases with presumptive tubercular lymphadenitis was 119. Majority of the aspirates are from jugular lymph nodal and cervical swellings 77 (64.7%) out of 119. FNAC has detected tuberculosis in 20 (16.8%) cases and ZN stain detected Acid Fast Bacilli (AFB) in 6 (5%) cases. CBNAAT has detected 28 (23.5%) cases, among them 15 (12.6%) cases which were not detected by FNAC. Diagnostic Performance of CBNAAT versus FNAC 28 out of 119 showed sensitivity, specificity, negative predictive value and positive predictive value 80%, 85.8%, 65% and 45.5%. Conclusion: The CBNAAT can be added with FNAC to get more specific results. CBNAAT is less sensitive for blood stained samples than purulent samples and hence FNAC still remains as the cheapest and first line test to diagnose in cases suspected of tubercular lymphadenoapathy.

Cryptosporidium infection among hemodialysis patients attending to the dialysis center at Kosti Teaching Hospital, Sudan.

Cryptosporidiosis is an illness caused by a protozooan parasite Cryptosporidium. Cryptosporidium species are an opportunistic pathogens cause a diarrheal disease worldwide, and can be more severe in immunocompromized patients. Until now, a little data have been available on its prevalence rate among haemodialysis patients in Sudan. Therefore, this article was designed to examine the prevalence of Cryptosporidium among hemodialysis Sudanese patients attending hemodialysis center at Kosti Teaching Hospital. A case-control study including one-hundred and twelve hemodialysis patients between November 2016 and January 2017 have been conducted. For the control group, we include one-hundred and twelve normal population. A total of two-hundred and twenty-four stool samples were collected. The stool samples were processed and examined using the modified Ziehl-Neelsen (ZN) staining method. High Cryptosporidium prevalence of 14/112 (12.5%) was detected in hemodialysis patients compare to the normal individuals 3/112 (2.7%). There was no correlation between the prevalence of Cryptosporidium infection with the age, sex, and the duration of dialysis (P>0.05). Therefore, an early detection and prompt treatment of Cryptosporidium infected hemodialysis patients is crucial.

Intestinal parasite infections in diabetes mellitus patients; A cross-sectional study of the Bolgatanga municipality, Ghana

Diabetes mellitus (DM) is a chronic disease which predisposes affected people to the acquisition of intestinal parasites. This study was conducted to detect the prevalence of intestinal parasites (ITPs) infection among DM patients and to identify their socio-demographic and clinical characteristics. The cross-sectional study was conducted among 152 diabetes mellitus patients selected from three hospitals in the Bolgatanga municipality. Stool samples were collected from the participants and examined using direct wet mount, formol-ether concentration, and modified Ziehl-Neelsen (ZN) staining methods. Questionnaires were designed to elicit socio-demographic information from participants and medical history was obtained from hospital folders. A prevalence of 12.5% ITP infections was recorded. Ascaris lumbricoides (5.3%), Cryptosporidium parvum (15%), Entamoeba hystolytica (21.1%), Entamoeba coli (15.8%), Giardia lamblia (47. 4%) and Hookworm (5.1%) were detected. Fasting blood glucose (FBG) (p<0.0001), duration of diabetes mellitus (p = 0.017), duration of therapy (p = 0.029) and type of therapy (metformin) (p = 0.0101) significantly affected the prevalence of ITPs infections. Diabetic mellitus complications (nephropathy, OR=25.20; CI=2.46–257.87; p<0.0001), no history of visit to dietician (OR=11.56; CI=3.95–33.82; p<0.0001) and level of education (informal education, OR=10.60; CI=1.22–92.27; p = 0.032) were significant risk factors of ITP infections. The Widowed (OR=0.13; CI=0.02–0.92; p = 0.040), married participants (OR=0.09; CI=0.02–0.35; p = 0.001), people that defecated twice a day (p<0.0001) and participants reported to have GI symptoms (p<0.0001) were also significantly associated with ITP infection. DM patients might be at an increased risk of infection with intestinal parasites especially G. lamblia, Entamoeba hystolytica and Cryptosporidium parvum and it is therefore necessary to carry out routine stool examination for them

Isolation and Genetic Characterization of Cryptosporidium from Captive Wildlife of India

Background: Cryptosporidiosis is an emerging zoonotic protozoan disease caused by Cryptosporidium spp. The infection was reported worldwide from domestic animals and humans, including wild animals. From India, no such reports were published on Cryptosporidium infection in captive wildlife. Hence, a pilot study was conducted to report the occurrence of Cryptosporidium infection in captive wildlife of India. Methods: Faecal samples (n=788) were collected from 127 captive wildlife species of three zoological parks viz., Sri Venkateswara Zoological Park (SVZP), Tirupati (n=242); Indira Gandhi Zoological Park (IGZP), Visakhapatnam (n=218); Nehru Zoological Park (NZP), Hyderabad (n=328) and screened for Cryptosporidium infection. Preliminary screening of faecal samples was done for the detection of Cryptosporidium oocysts by modified Ziehl-Neelsen (mZN) staining method and the test positives were confirmed by nested PCR targeting 18S rRNA gene. Nested PCR amplicons were sequenced for determining the Cryptosporidium species. The resultant data were statistically analyzed by Fisher/Chi square, Fisher Exact test using SPSS software v 17.0.Result: In mZN staining method, 7.23 percent of isolates were found to be positive for Cryptosporidium and the highest rate of infection was detected in wildlife at NZP, Hyderabad (8.23%), followed by SVZP, Tirupati (7.44%) and IGZP, Visakhapatnam (5.50%). Cryptosporidium positive faecal samples by mZN staining were further confirmed by nested PCR and positive amplicons were sequenced for determination of Cryptosporidium species. Genetic characterization revealed five species viz., Cryptosporidium parvum; C. ryanae, C. suis, C. muris and Cryptosporidium avian genotype III. The study conclude that, Cryptosporidium infection was prevalent in the captive wildlife from the zoological parks of India and species variation was marked among the wildlife. Based on the available literature, the current study is the first of its kind on the prevalence of Cryptosporidium in captive wildlife from India.

Assessment of Cryptosporidium in Patients With Gastroenteritis by Modified Ziehl-Neelsen Staining Method in East Azerbaijan Province of Iran During 2018-2019

Introduction: Cryptosporidiosis is a worldwide zoonotic disease that is caused by Cryptosporidium species and leads to acute or chronic diarrhea and vomiting in patients. Due to the suitable conditions of East Azerbaijan province for parasite transmission, the present study was performed to evaluate the prevalence of Cryptosporidium infection in patients with gastroenteritis referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. Methods: A descriptive cross-sectional study was performed on patients with diarrhea who referred to hospitals and reference laboratories of East Azerbaijan province during 2018-2019. In this study, 180 fecal samples were collected from patients with gastroenteritis by cluster-random sampling. Stool samples were stored in 10% formalin and transferred to a parasitology laboratory and examined for parasite contamination by modified acid-fast staining (Modified Ziehl-Neelsen) method. Data were analyzed by chi-square test using SPSS. Results: Out of 180 samples, the highest number of diarrhea samples belonged to the age group over 40 years (35.5%) and the lowest belonged to the age group of 10-20 years (7.7%). Additionally, 56% of subjects were male and 44% were female. Of these patients, 57.7% lived in urban areas and 42.3% in rural areas. Cryptosporidium was observed in 1.6% (3 cases) of patients with gastroenteritis. There was a statistically significant relationship between Cryptosporidium infection and the age of patients with diarrhea (P&lt;0.05). Conclusion: In the present study, a lower prevalence of cryptosporidiosis has been determined in comparison with previous studies in East Azerbaijan province. However, Cryptosporidium and other parasitic infections in the feces of patients with diarrhea referred to hospitals or reference laboratories need to be carefully diagnosed by appropriate parasitological methods.